Swiss Institute for Experimental Cancer Research (ISREC), School of Life Sciences, Swiss Federal Institute of Technology (EPFL), 1015 Lausanne, Switzerland.
Dev Cell. 2013 Jun 24;25(6):555-71. doi: 10.1016/j.devcel.2013.05.016. Epub 2013 Jun 13.
Centrioles are essential for forming cilia, flagella, and centrosomes and are thus critical for a range of fundamental cellular processes. Despite their importance, the mechanisms governing centriole biogenesis remain incompletely understood. We performed a high-content genome-wide small-interfering-RNA-based screen to identify genes regulating centriole formation in human cells. We designed an algorithm to automatically detect GFP-Centrin foci that, combined with subsequent manual analysis, allowed us to identify 44 genes required for centriole formation and 32 genes needed for restricting centriole number. Detailed follow-up characterization uncovered that the C2 domain protein C2CD3 is required for distal centriole formation and suggests that it functions in the basal body to template primary cilia. Moreover, we found that the E3 ubiquitin ligase TRIM37 prevents centriole reduplication events. We developed a dynamic web interface containing all images and numerical features as a powerful resource to investigate facets of centrosome biology.
中心体对于形成纤毛、鞭毛和中心体至关重要，因此对于一系列基本的细胞过程至关重要。尽管它们很重要，但控制中心体生物发生的机制仍不完全清楚。我们进行了高通量全基因组小干扰 RNA 为基础的筛选，以鉴定在人类细胞中调节中心体形成的基因。我们设计了一种自动检测 GFP-Centrin 焦点的算法，该算法与后续的手动分析相结合，使我们能够识别出 44 个中心体形成所必需的基因和 32 个限制中心体数量所必需的基因。详细的后续特征表明，C2 结构域蛋白 C2CD3 是远端中心体形成所必需的，并表明它在基体中起模板初级纤毛的作用。此外，我们发现 E3 泛素连接酶 TRIM37 可防止中心体重复事件。我们开发了一个动态的网络界面，其中包含所有的图像和数值特征，作为一个强大的资源，以研究中心体生物学的各个方面。
